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EVALUATION OF THE PULSED

DISCHARGE HELIUM IONIZATION


DETECTOR
FOR THE ANALYSIS OF
HYDROGEN AND METHANE IN
BREATH
1.
2.
3.
4.
5.

Nama anggota:
Kartika Nur A (191339
Purwanita Indah K (19133966A)
Erni Sukmawati K (19133967A)
Yulinda Kussukmawaty(19133972A)
Wisnu Daelani (19

ABSTRACT
Under the appropriate separation conditions the pulsed discharge
helium ionization detector (PDHID) was used to detect hydrogen
and methane separated from the matrix components of human
breath samples. The sensitivity of this method is over an order of
magnitude better than published methods using a flame ionization
detector (FID) and a thermal conductivity detector (TCD), and has
the further advantage of detecting both analytes with only one
detector. Limits of detection were 0.3 ppmv for both hydrogen and
methane and the method had a linear dynamic range (LDR) of three
orders of magnitude (0.3400 ppm, v/v). The PDHID was also
compared to the FID and the TCD in regard to selectivity, sensitivity
and reproducibility for high-speed gas chromatography (HSGC). It
was shown that the PDHID is as sensitive as the FID for fast
separations but is limited by the difficulty of resolving analyte
peaks from O2 and N2. The PDHID was at least three orders of
magnitude more sensitive than the TCD for all of the analytes
examined

MATERIALS AND METHODS


Instrumentation
1.Sampling and injection
Gaseous samples were injected using a loop injector. The loop injector
consisted of a Valco six-port, two-position valve (Valco, Houston, TX). The
sample loop (9 l), as well as all tubing and connections, were 1/16 in.
stainless steel. Samples were pulled through the sampling loop using a
Gast model DOA-P104-AA rotary pump (Daigger, Vernon Hills, IL, USA).
2. Gas chromatographs
GCPDHID experiments were conducted either with a Hewlett-Packard
5890 oven or with an oven constructed in our laboratory. In each case, a
Valco model D-1 PDHID and electrometer were installed (Valco, Houston,
TX, USA). Data obtained from both instruments were nearly identical. All
data collection occurred on a Hewlett-Packard HP 3396 Series II integrator
(Hewlett-Packard,Wilmington, DE, USA). UHP helium (Airgas) was used as
both the carrier and discharge gas. An Indicating Oxytrap (Alltech,
Deerfield, IL, USA) followed by a Valco model HP helium purifier (Valco)
were placed between the cylinder and the flow splitter.

A Hewlett-Packard 5890A GC with a TCD using UHP


helium carrier gas was used for comparison and was
configured using the six-port loop injector described for
the PDHID system.
3. Separation conditions
H2, O2, N2, CH4 and CO were separated on a molecular sieve
5A porous-layer open tubular (PLOT) column, 30m 0.32 mm,
held at 35 C (Supelco, Bellefonte, PA, USA). Flow restrictors
were used to set the carrier gas flow rate at 5.2 ml/min for
comparisons of the PDHID to the TCD. The GCTCD used a carrier
gas flow rate of 2 ml/min. Make up gas was added to reach the
recommended 5 ml/min and the reference gas was set at 20
ml/min. The injector, oven and detector temperatures were set at
100, 80 and 200 C, respectively.

Msieve 5A column (Restek, Bellefonte, PA, USA). Flow was set


at 2 ml/min using flow restrictors. The oven temperature was
30oC while the detector was 190 C.
4. Human breath samples
The effects of diet on colonic fermentation was assessed by
measuring hydrogen and methane production. Samples were
collected from non-smoking males ages 36 and 41 years with
body mass indexes (BMI) of 28.5 and 34.5, respectively. Both
subjects were screened for general health before they were
allowed into the study. Subjects gave their informed consent to
participate in the study and all aspects of the study were
reviewed and approved by the Institutional Review Board of the
University of North Dakota.
Hydrogen and methane were determined from expired air
samples collected before and after exercise-induced oxidative
stress. Prolonged exercise performance tests were conducted
using a stationary cycle (ergocycle). Subjects pedaled the
ergocycle for 45 min at 70% of maximum heart rate, and 5 min
at 90%. Breath samples were collected before and 7 min after

Samples were collected using a one-way breathing


valve (Hans Rudolph, Kansas City, MO, USA) connected
to 20 l Cali-5-Bond sampling bags (Calibrated
Instruments, Hawthorne, NY, USA) and analyzed within
3 h of collection. The one-way breathing
valve was attached to the two-way valve of the Cali-5Bond
sampling bag. Subjects were seated and instructed to
breath
naturally until their expired breath completely filled the
sample bag. Two-way valves were opened immediately
prior to the subjects first exhalation and closed
immediately after their final exhalation. Calibration
curves were used to quantitate analytes in samples.

RESULTS AND DISCUSSION


PDHID versus TCD selectivity and sensitivity
Both the PDHID and the TCD are essentially nonselective detectors, and either could be used to measure
both hydrogen and methane in breath samples. One
advantage of the PDHID is in its greater sensitivity to light
gases. As expected, the TCD showed little or no response
to hydrogen, which does not have a significantly different
thermal Conductivity than the helium carrier gas, while
the PDHID was useful for this measurement. Because both
the PDHID and the TCD are universal for all other
analytes, both are equally affected by the potential
interferences from the air matrix.

Limits of detection for the TCD appear rather high,


but they are based on the small injection volumes (9 l)
used in these experiments. Small volume loops help to
minimize injection band duration and allow high-speed
separations to be made with good resolution. Limits of
detection for the TCD have been reported to be 106 to
108 g, which are in agreement with our values at these
injection volumes. In comparison, the total minimum
quantity detected by the PDHID in our experiments was
1010 to 1012 g which is similar to results published
by Wentworth et al. The more sensitive PDHID can detect
analytes at levels in the low ppmv range under fast
separation conditions using a small volume injection
loop, which is not possible with the TCD. This is greatly
advantageous for our studies monitoring hydrogen and
methane
in
breath
samples.

Application to human breath


It has been reported that approximately half of the human
population emits hydrogen and methane in their breath
as a product of microbial fermentation in the colon.
In the past, separations of human breath were detected
using the TCD for hydrogen and an FID for methane,
with both methods having a limit of detection of 6 ppmv. As
mentioned above, the TCD has poor sensitivity
for hydrogen. We were able to use the universal PDHID
as a detector for both analytes by using a small volume
injection on a capillary column to separate methane from
nitrogen.

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